Effects of vibration on the expression of mitochondrial fusion and fission genes and ultrastructure of skeletal muscle in rabbits / 中华劳动卫生职业病杂志

Objective: To study the effects of vibration on the expression of mitochondrial fusion and fission genes and ultrastructure of skeletal muscle in rabbits. Methods: Thirty-two 3.5-month-old New Zealand rabbits were randomly divided into low-intensity group, medium-intensity group, high-intensity group and control group, with 8 rabbits in each group. The rabbits in the experimental group were subjected to hind limb vibration load test for 45 days. The vibration intensity of the high intensity group was 12.26 m/s(2), the medium intensity group was 6.13 m/s(2), and the low intensity group was 3.02 m/s(2) according to the effective value of weighted acceleration[a(hw (4))] for 4 hours of equal energy frequency. The control group was exposed to noise only in the same experimental environment as the medium-intensity group. The noise levels of each group were measured during the vibration load experiment. After the test, the mRNA expression of mitochondrial fusion gene (Mfn1/Mfn2) and fission gene (Fis1, Drp1) by RT-PCR in the skeletal muscles were measured and the ultrastructure of the skeletal muscles were observed in high intensity group. Results: The mRNA expression of mitochondrial in the skeletal muscle tissues of control group, low intensity group, medium intensity group and high intensity group were Mfn1: 3.25±1.36, 3.85±1.90, 4.53±2.31 and 11.63±7.68; Mfn2: 0.68±0.25, 1.02±0.40, 0.94±0.33 and 1.40±0.45; Fis1: 1.05±0.62, 1.15±0.59, 1.53±1.06 and 2.46±1.51 and Drp1: 3.72±1.76, 2.91±1.63, 3.27±2.01 and 4.21±2.46, respectively. Compared with the control group, the expressions of Mfn1 mRNA, Mfn2 mRNA and Fis1 mRNA in the high-intensity group increased significantly (P<0.05) , and the expressions of Mfn2 mRNA in the medium-intensity group and the low-intensity group increased significantly (P<0.05) . Compared with the control group, the ultrastructure of skeletal muscle of high intensity group showed mitochondrial focal accumulation, cristae membrane damage, vacuole-like changes; Z-line irregularity of muscle fibers, and deficiency of sarcomere. Conclusion: Vibration must be lead to the abnormal mitochondrial morphology and structure and the disorder of energy metabolism due to the expression imbalance of mitochondrial fusion and fission genes in skeletal muscles of rabbits, which may be an important target of vibration-induced skeletal muscle injury.

Clinical and morphological justification of the treatment of purulonecrotic lesion in deep structures of limb distal segment in pig: case report / [Justificativa clínica e morfológica do tratamento de lesão purulonecrótica em estruturas profundas do segmento distal de membros em porco: relato de caso]

Surgical pathology of the musculoskeletal system, and in particular the diseases of the limb distal segment in pigs are quite common. Their significant spread leads to economic losses due to culling, compulsory slaughter, short-received pig production and pig crop. The purpose of this work was to reveal clinical and morphological features of limb distal segment diseases in pigs and to study the possibility of preserving their health and productivity. The conservative treatment of purulonecrotic lesions in the deep structures of the limb distal segment in pigs is not promising. It is shown that the best way to treat a given pathology is amputation of a sick limb. The technique of carrying out exarticulation of talus shin consists in separation of soft tissues, capsule and ligaments, ligation of vessels, formation of stump. The postoperative recovery period of the animal body is 25 days.(AU)

A patologia cirúrgica do sistema musculoesquelético e, em particular, as doenças do segmento distal dos membros em suínos são bastante comuns. A sua propagação significativa leva a perdas econômicas devido ao abate seletivo, abate obrigatório, produção de suínos pouco recebida e colheita de suínos. O objetivo deste trabalho foi revelar características clínicas e morfológicas das doenças do segmento distal de membros em suínos e estudar a possibilidade de preservar sua saúde e produtividade. O tratamento conservador das lesões purulonecróticas nas estruturas profundas do segmento distal do membro em porcos não é promissor. Fica estabelecido que a melhor forma de tratar uma determinada patologia é a amputação de um membro doente. A técnica de realizar a exarticulação da canela do talos consiste na separação dos tecidos moles, cápsula e ligamentos, ligadura dos vasos, formação do coto. O período de recuperação pós-operatória do corpo do animal é de 25 dias.(AU)

An atypical presentation of leiomyosarcoma causing extremity compartment syndrome of the crural region in a Dutch Warmblood mare: a case report

Exploring the role of neurogenic pathway-linked cholecystokinin release in remote preconditioning-induced cardioprotection

Abstract Purpose: The current study explored the involvement of neurogenic pathway-linked cholecystokinin (CCK) release in RIP-induced cardioprotection in rats. Methods: Male Wistar rats were subjected to four cycles of alternate episodes of ischemia and reperfusion (five min each) to induce RIP. Thereafter, the hearts were subjected to global ischemia and reperfusion ex vivo. The myocardial damage was assessed by quantifying the levels of heartspecific biochemicals i.e. LDH-1, CK-MB and cTnT. Apoptotic cell injury was assessed by measuring the levels of caspase-3 and Bcl-2. The levels of CCK were measured in the plasma following RIP. Results: Exposure to RIP significantly increased the plasma levels of CCK and attenuated IR-induced myocardial injury. Administration of CCK antagonist, proglumide significantly attenuated RIP-induced cardioprotection. Administration of hexamethonium, a ganglion blocker, abolished RIP-induced increase in plasma CCK levels and cardioprotective effects. Exogenous delivery of CCK-8 restored the effects of RIP in hexamethonium treated animals. Conclusion: RIP activates the neurogenic pathway that may increase the plasma levels of CCK, which may act on the heart-localized CCK receptors to produce cardioprotection against I/R injury.

Anatomia óssea e muscular do antebraço e mão de Chrysocyon brachyurus (Carnivora, Canidae) / Bone and muscular anatomy of the antibone and handof Chrysocyon brachyurus (Carnivora, Canidae)

O lobo-guará Chrysocyon brachyurus Illiger, 1815, é o maior canídeo da América do Sul, pesa cerca de 25 kg quando adulto e está ameaçado de extinção. Descrições anatômicas contribuem para a complementação das informações sobre espécies silvestres e para implicações conservacionistas, clínicas e cirúrgicas. Objetivou-se descrever os ossos e os músculos do antebraço e mão do lobo-guará. A preparação das peças foi feita a partir dos métodos usuais de dissecação, em animais preservados em solução de formol a 10%. Os espécimes pertencem ao acervo didático do Laboratório de Ensino e Pesquisa em Animais Silvestres da UFU e são provenientes de indivíduos atropelados. Os ossos descritos foram: rádio, ulna, ossos cárpico acessório, cárpico ulnar e cárpico intermédio; ossos cárpicos I, II, III e IV; ossos metacárpicos I, II, III, IV, V; falanges proximais, falanges médias e falanges distais do primeiro ao quinto dedo. Os músculos observados foram: extensor radial do carpo; pronador redondo; braquiorradial; extensor comum dos dedos; extensor ulnar do carpo; extensor lateral dos dedos; supinador; abdutor longo do dedo I; flexor radial do carpo; flexor profundo dos dedos; flexor superficial dos dedos; flexor ulnar do carpo; pronador quadrado; interflexor; lumbricais; abdutor curto dos dedos I e II e flexor curto do dedo I.

The maned wolf Chrysocyon brachyurus (Illiger, 1815) is the largest canid in South America, weighs about 25 kg as an adult and is threatened of extinction. Anatomical descriptions contribute to the complementation of information on wild species and for conservation, clinical and surgical implications. The purpose of this study was to describe the bones and muscles of the forearm and hand of the maned wolf. The methodology was through the usual dissecting methods in animals preserved in 10% formalin solution. The animals belong to the didactic collection of the Laboratory of Teaching and Research in Wild Animals of the UFU and come from run over. The bones evaluated were: radius, ulna, carpal accessory, carpi ulnar and carpi intermedium; carpal bones I, II, III and IV; metacarpal bones I, II, III, IV, V; proximal phalanges, middle phalanges and distal phalanges from first to fifth finger. The muscles observed were: radial extensor carpal; pronator round; brachioradial; common extensor of fingers; ulnar carpal extensor; lateral extensor of the fingers; supinator; abductor long finger I; flexor carpi radialis; flexor deep fingers; superficial flexor of the fingers; ulnar flexor of the carpus; square pronator; interflexor; lumbrils; short abductor of fingers I and II and short flexor of finger I.

Preoperative anxiety induces chronic postoperative pain by activating astrocytes in the anterior cingulate cortex region

SUMMARY OBJECTIVE The study aims to explore the relationship between preoperative anxiety and chronic postoperative pain. METHODS A total of forty rats were divided into four groups, control, single-prolonged stress alone, Hysterectomy alone, and SPS+ Hysterectomy. The paw withdrawal mechanical thresholds (PWMT) were examined. qRT-PCR and western blotting assay were performed to detect the GFAP expression in astrocytes isolated from the anterior cingulate cortex (ACC) region. In addition, the long-term potentiation (LTP) in ACC was examined. RESULTS Rats in the SPS group or the Hysterectomy alone group had no significant effect on chronic pain formation, but SPS can significantly induce chronic pain after surgery. Astrocytes were still active, and the LTP was significantly increased three days after modeling in the SPS+Hysterectomy group. CONCLUSIONS anxiety can induce chronic pain by activating astrocytes in the ACC region.

RESUMO OBJETIVO O objetivo deste estudo é explorar a relação entre a ansiedade no pré-operatório e a dor crônica no pós-operatório. MÉTODOS Um total de 40 ratos foram divididos em quatro grupos: controle, estresse prolongado (SPS), histerectomia e SPS + histerectomia. Os limiares de retirada da pata em resposta a estímulo mecânico (PWMT) foram examinados. Ensaios qRT-PCR e imunoenzimáticos (western blotting) foram realizados para detectar a expressão de GFAP em astrócitos isolados da região do córtex cingulado anterior (CCA). Além disso, a potenciação de longa duração (LTP) no CCA também foi examinada. RESULTADOS Os ratos no grupo de estresse prolongado e no grupo de histerectomia não apresentaram nenhum efeito significativo na formação de dor crônica. Porém, o estresse prolongado foi capaz de induzir dor crônica significativamente após a cirurgia. Três dias após o modelo, o grupo de SPS + histerectomia ainda apresentava astrócitos ativos e LTP significativamente maior. CONCLUSÃO A ansiedade pode provocar dor crônica através da ativação de astrócitos na região do CCA.

Are cursorial birds good kinematic models of non-avian theropods? / ¿Son los pájaros corredores buenos modelos cinemáticos de terópodos no aviares?

Determining kinematics of hindlimbs of theropod dinosaurs has been a challenge. Since cursorial birds are phylogenetically closest to theropod dinosaurs they are commonly used as a kinematic model of theropod dinosaur locomotion. Using a comparative biomechanical approach, we found that cursorial birds have a different morphology of legs than non avian theropodos and that appears to be that felines and ungulates share more morphological properties in the hindlimbs with theropod dinosaurs than cursorial birds. We calculated the ratio between the lower leg and the femur, and the relative length of the tibia and the metatarsus with respect to the length of the femur in cursorial birds, as well as felines, ungulates and non-avian theropods. We found that as the length of the femur increases, the length of the lower leg increases similarly in felines, ungulates and non-avian theropods. On the other hand, existing and extinct cursorial birds did not follow this pattern. This observation suggests that the hindlimb of cursorial birds are not well suited to serve as kinematic models for hindlimb of extinct theropod dinosaur locomotion.

Determinar la cinemática de los miembros pelvianos de los dinosaurios terópodos ha sido un desafío. Dado que las aves corredoras son filogenéticamente más cercanas a los dinosaurios terópodos, son comúnmente utilizadas como modelo cinemático de la locomoción del dinosaurio terópodo. Usando un enfoque biomecánico comparativo, encontramos que las aves corredoras tienen una morfología de pies diferente a la de los terópodos no aviares y parece ser que los felinos y los ungulados comparten más propiedades morfológicas en los pies con los dinosaurios terópodos que las aves corredoras. Calculamos la proporción entre la parte inferior de la pierna y el fémur, y la longitud relativa de la tibia y el metatarso con respecto a la longitud del fémur en aves corredoras, así como en los terópodos no aviares y ungulados. Encontramos que a medida que aumenta la longitud del fémur, la longitud de la parte inferior de la pierna aumenta de manera similar en los terópodos, los ungulados y los terópodos no aviares. Por otro lado, las aves corredoras existentes y extintas no siguieron este patrón. Esta observación sugiere que el miembro pelviano de las aves corredoras no es adecuada para servir como modelos cinemáticos de locomoción del miembro pelviano del dinosaurio terópodo extinto.

Murine Model Study of a New Receptor-Targeted Tracer for Sentinel Lymph Node in Breast Cancer / 한국유방암학회지

PURPOSE: Sentinel lymph node biopsy (SLNB), a critical staging and treatment step, has replaced axillary lymph node (LN) dissection as the standard staging procedure for early stage breast cancer patients with clinically negative axillary LNs. Hence, using a murine sentinel lymph node (SLN) model, we investigated the localization effect of the new receptor-targeted tracer, indocyanine green (ICG)-rituximab, on breast cancer SLNB. METHODS: After establishing the murine SLN model, different doses of ICG-rituximab were subcutaneously injected into the hind insteps of BALB/c mice to determine the optimal dose and imaging time using continuous (> 3 hours) MDM-I fluorescence vasculature imaging. To explore the capacity of ICG-rituximab for sustained SLN localization with the optimal dose, MDM-I imaging was monitored at 6, 12, and 24 hours. RESULTS: The popliteal LN was defined as the SLN for hindlimb lymphatic drainage, the iliac LN as the secondary, and the para-aortic or renal LN as the tertiary LNs. The SLN initial imaging and optimal imaging times were shortened with increased ICG-rituximab doses, and the imaging rates of the secondary and tertiary LNs increased accordingly. The optimal ICG dose was 0.12 μg, and its optimal imaging time was 34 minutes. After 24 hours, the SLN imaging rate remained 100%, while those of the secondary and the tertiary LNs increased from 0% (6 hours) and 0% (6 hours) to 10% (12 hours) and 10% (12 hours) to 20% (24 hours) and 10% (24 hours), respectively. CONCLUSION: ICG-rituximab localized to the SLN without imaging from the secondary or tertiary LNs within 6 hours. The optimal ICG dose was 0.12 μg, and the optimal interval for SLN detection was 34 minutes to 6 hours post-injection. This novel receptor-targeted tracer is of great value to clinical research and application.

Antiedematogenic activity of the indole derivative N-salicyloyltryptamine in animal models

ABSTRACT The N-salicyloyltryptamine (NST) is an indole derivative compound analogue to the alkaloid N-benzoyltryptamine. In the present study, the antiedematogenic activity of NST was investigated in animal models. Firstly, the acute toxicity for NST was assessed according to the OECD Guideline no. 423. The potential NST-induced antiedematogenic activity was evaluated by carrageenan-induced paw edema in rats, as well as by dextran-, compound 48/80-, histamine-, serotonin-, capsaicine-, and prostaglandin E2-induced paw edema in mice. The effect of NST on compound 48/80-induced ex vivo mast cell degranulation on mice mesenteric bed was investigated. No death or alteration of behavioral parameters was observed after administration of NST (2000 mg/kg, i.p.) during the observation time of 14 days. The NST (100 and 200 mg/kg, i.p.) inhibited the carrageenan-induced edema from the 1st to the 5th hour (**p<0.01; ***p<0.001). The edematogenic activity induced by dextran, compound 48/80, histamine, serotonin, capsaicin, and prostaglandin E2 was inhibited by NST (100 mg/kg, i.p.) throughout the observation period (**p<0.01; ***p<0.001). The pretreatment with NST (50, 100 or 200 mg/kg, i.p) attenuates the compound 48/80-induced mast cell degranulation (**p<0.01; ***p<0.001). Thus, the inhibition of both mast cell degranulation and release of endogenous mediators are probably involved in the NST-induced antiedematogenic effect.

Genetic Ablation of EWS RNA Binding Protein 1 (EWSR1) Leads to Neuroanatomical Changes and Motor Dysfunction in Mice

A recent study reveals that missense mutations of EWSR1 are associated with neurodegenerative disorders such as amyotrophic lateral sclerosis, but the function of wild-type (WT) EWSR1 in the central nervous system (CNS) is not known yet. Herein, we investigated the neuroanatomical and motor function changes in Ewsr1 knock out (KO) mice. First, we quantified neuronal nucleus size in the motor cortex, dorsal striatum and hippocampus of three different groups: WT, heterozygous Ewsr1 KO (+/−), and homozygous Ewsr1 KO (−/−) mice. The neuronal nucleus size was significantly smaller in the motor cortex and striatum of homozygous Ewsr1 KO (−/−) mice than that of WT. In addition, in the hippocampus, the neuronal nucleus size was significantly smaller in both heterozygous Ewsr1 KO (+/−) and homozygous Ewsr1 KO (−/−) mice. We then assessed motor function of Ewsr1 KO (−/−) and WT mice by a tail suspension test. Both forelimb and hindlimb movements were significantly increased in Ewsr1 KO (−/−) mice. Lastly, we performed immunohistochemistry to examine the expression of TH, DARPP-32, and phosphorylated (p)-DARPP-32 (Thr75) in the striatum and substantia nigra, which are associated with dopaminergic signaling. The immunoreactivity of TH and DARPP-32 was decreased in Ewsr1 KO (−/−) mice. Together, our results suggest that EWSR1 plays a significant role in neuronal morphology, dopaminergic signaling pathways, and motor function in the CNS of mice.

Microvascular in-vivo analysis of retrograde venous arterialization of ischemic skeletal muscle

@#<p style="text-align: justify;"><strong>OBJECTIVE:</strong> Nearly 20% of patients with critical limb ischemia will not be suitable for arterial bypass due to distal small vessel occlusion, and venous arterialization of the distal venous bed might be a valuable surgical option. This study demonstrates the in-vivo microcirculatory effects of this type of intervention.</p><p style="text-align: justify;"><strong>METHODS:</strong> Using intravital video microscopy, the authors studied the distal skeletal microcirculatory characteristics following venous arterialization of critical hindlimb ischemia in the rat. 25 Wistar rats underwent proximal ligation of the femoral arteries followed by venous arterialization carried out by anastomosing the saphenous vein to the femoral artery using microsurgery techniques. Microcirculatory hemodynamic conditions of the soleus muscle were observed under normal, ischemic, and arterialized conditions. Fluorescein-labeled red cells were used to measure red cell velocities (Vrbc) at the capillaries, and acridine orange injections used to stain endothelial cell nuclei to measure microcirculatory diameters, and leukocyte nuclei to measure leukocyte adhesion. Laser Doppler Perfusion (LDP) units at the distal limb were measured continuously throughout the procedure.</p><p style="text-align: justify;"><strong>RESULTS:</strong> Proximal femoral arterial ligation resulted in drastic reductions in LDP and Vrbc. Following distal venous arterialization, LDP returned to an average of 41% of baseline. Vrbc returned to near baseline values in 70% of the capillaries. Flow at the capillary and venular system showed frequent reversals and great variations in velocities. Venules and venu-venular anastomoses diameters increased by 50%. There was immediate macromolecular tracer leakage and leukocyte activation was significantly increased in both ischemic and arterialized groups (15 cells vs 156 and 178 cells respectively).</p><p style="text-align: justify;"><strong>CONCLUSION:</strong> Venous arterialization may provide an improvement in microcirculatory velocities but is accompanied by microcirculatory injury and dysfunction in the acute phase. These results suggest that mechanisms besides microcirculatory hemodynamics play a role in the overall picture of clinical effectivity of the procedure</p>

Proposal for a new histological scoring system for cartilage repair

OBJECTIVE: This study aimed to develop a new histological scoring system for use in a partial-thickness cartilage repair animal model. Although previous papers have investigated the regeneration of articular cartilage, the good results achieved in small animals have not been replicated in large animal models or humans, possibly because of the frequent use of models with perforation of the subchondral bone plates. Partial-thickness lesions spare the subchondral bone, and this pattern is the most frequent in humans; therefore, new therapies should be tested using this model. However, no specific histological score exists to evaluate partial-thickness model results. METHODS: Histological sections from 30 ovine knees were reviewed to develop a new scoring system. The sections were subjected to H&E, Safranin O, and Masson's trichrome staining. RESULTS: This paper describes a new scoring tool that is divided into sections in detail: repair of tissue inside the lesion, cartilage around the lesion and degenerative changes at the base of the lesion. Scores range from 0 to 21; a higher score indicates better cartilage repair. DISCUSSION: Unlike existing tools, this new scale does not assign points for the positioning of a tidemark; we propose evaluation of the degenerative changes to the subchondral bone and calcified cartilage layer. It is necessary to remove the whole joint to access and study the evolution of the lesion as well as the surrounding tissue. CONCLUSION: This article emphasizes the importance of a partial-thickness animal model of cartilage repair and presents a new histological scoring system.

Transferência de autoenxerto osteocondral para tratamento de osteocondrite dissecante em côndilo femoral de cão / Osteochondral autograft transfer for treatment of osteochondritis dissecans of the femoral condyle in a dog

Na osteocondrite dissecante (OCD), a articulação do ombro é mais comumente afetada, mas o joelho é ocasionalmente lesionado, o que, muitas vezes, passa desapercebido. O tratamento cirúrgico precoce é indicado para remover cartilagem solta, aliviar a dor e minimizar a artrose. Benefícios putativos da transferência de autoenxerto osteocondral em relação às técnicas convencionais incluem a reconstrução exata do contorno subcondral e articular, recobrimento da superfície com cartilagem hialina e criação de uma barreira imediata entre o líquido sinovial e o osso subcondral. O objetivo deste trabalho é relatar a técnica de transferência de autoenxerto osteocondral para o tratamento da osteocondrite do côndilo femoral. Um cão da raça Bull Terrier foi tratado cirurgicamente por meio da técnica de transferência de autoenxerto osteocondral, após ter sido diagnosticado com OCD do côndilo femoral, apresentando melhora clínica significativa e completa recuperação aos 30 dias de pós-operatório.(AU)

In osteochondritis dissecans (OCD), the shoulder joint is most commonly involved, but the stifle is occasionally involved and often goes unnoticed. Early surgical treatment is indicated to remove loose cartilage and minimize osteoartrosis. Putative benefits of Osteochondral Autograft Transfer (OAT) over conventional techniques include accurate reconstruction of subchondral and articular contour, resurfacing with hyaline or hyaline-like cartilage, and creation of an immediate barrier between synovial fluid and subchondral bone. The purpose of this work is to report the technique of OATs for the treatment of osteochondritis of the femoral condyle. A canine attended the Laboratory of Comparative Orthopedics and Traumatology USP-FMVZ underwent surgery after being diagnosed with OCD of the femoral condyle, with significant clinical improvement at 30 days postoperatively.(AU)

Rapid generation of OPC-like cells from human pluripotent stem cells for treating spinal cord injury

Remyelination via the transplantation of oligodendrocyte precursor cells (OPCs) has been considered as a strategy to improve the locomotor deficits caused by traumatic spinal cord injury (SCI). To date, enormous efforts have been made to derive OPCs from human pluripotent stem cells (hPSCs), and significant progress in the transplantation of such cells in SCI animal models has been reported. The current methods generally require a long period of time (>2 months) to obtain transplantable OPCs, which hampers their clinical utility for patients with SCI. Here we demonstrate a rapid and efficient method to differentiate hPSCs into neural progenitors that retain the features of OPCs (referred to as OPC-like cells). We used cell sorting to select A2B5-positive cells from hPSC-derived neural rosettes and cultured the selected cells in the presence of signaling cues, including sonic hedgehog, PDGF and insulin-like growth factor-1. This method robustly generated neural cells positive for platelet-derived growth factor receptor-α (PDGFRα) and NG2 (~90%) after 4 weeks of differentiation. Behavioral tests revealed that the transplantation of the OPC-like cells into the spinal cords of rats with contusive SCI at the thoracic level significantly improved hindlimb locomotor function. Electrophysiological assessment revealed enhanced neural conduction through the injury site. Histological examination showed increased numbers of axon with myelination at the injury site and graft-derived myelin formation with no evidence of tumor formation. Our method provides a cell source from hPSCs that has the potential to recover motor function following SCI.

Remote Ischemic Conditioning by Effluent Collected from a Novel Isolated Hindlimb Model Reduces Infarct Size in an Isolated Heart Model

BACKGROUND AND OBJECTIVES: Experimental protocols for remote ischemic conditioning (RIC) utilize models in which a tourniquet is placed around the hindlimb or effluent is collected from an isolated heart. In analyzing the humoral factors that act as signal transducers in these models, sampled blood can be influenced by systemic responses, while the effluent from an isolated heart might differ from that of the hindlimb. Thus, we designed a new isolated hindlimb model for RIC and tested whether the effluent from this model could affect ischemia/reperfusion (IR) injury and if the reperfusion injury salvage kinase (RISK) and survivor activating factor enhancement (SAFE) pathways are involved in RIC. MATERIALS AND METHODS: After positioning needles into the right iliac artery and vein of rats, Krebs-Henseleit buffer was perfused using a Langendorff apparatus, and effluent was collected. The RIC protocol consisted of 3 cycles of IR for 5 minutes. In the RIC effluent group, collected effluent was perfused in an isolated heart for 10 minutes before initiating IR injury. RESULTS: Compared with the control group, the infarct area in the RIC effluent group was significantly smaller (31.2%±3.8% vs. 20.6%±1.8%, p<0.050), while phosphorylation of signal transducer and activation of transcription-3 (STAT-3) was significantly increased. However, there was a trend of increased phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 in this group. CONCLUSION: This is the first study to investigate the effect of effluent from a new isolated hindlimb model after RIC on IR injury in an isolated heart model. The RIC effluent was effective in reducing the IR injury, and the cardioprotective effect was associated with activation of the SAFE pathway.

Engineered M13 Nanofiber Accelerates Ischemic Neovascularization by Enhancing Endothelial Progenitor Cells

Dysfunction or loss of blood vessel causes several ischemic diseases. Although endothelial progenitor cells (EPCs) are a promising source for cell-based therapy, ischemia-induced pathophysiological condition limits the recovery rate by causing drastic cell death. To overcome this issue, we attempted to develop a cell-targeted peptide delivery and priming system to enhance EPCbased neovascularization using an engineered M13 bacteriophage harboring nanofibrous tubes displaying ∼ 2700 multiple functional motifs. The M13 nanofiber was modified by displaying RGD, which is an integrin-docking peptide, on the minor coat protein, and bymutilayering SDKPmotifs,which are the key active sites for thymosin b4, on themajor coat protein. The engineered M13 nanofiber dramatically enhanced ischemic neovascularization by activating intracellular and extracellular processes such as proliferation, migration, and tube formation in the EPCs. Furthermore, transplantation of the primed EPCs with the M13 nanofiber harboring RGD and SDKP facilitated functional recovery and neovascularization in a murine hindlimb ischemia model. Overall, this study demonstrates the effectiveness of theM13 nanofiber-based novel peptide deliveryandprimingstrategy inpromotingEPC bioactivity and neovessel regeneration. To our knowledge, this is first report onM13 nanofibers harboring dual functional motifs, the use of which might be a novel strategy for stem and progenitor cell therapy against cardiovascular ischemic diseases.

Comparison of the Skin and Skeletal Muscle Contusion in Rats Induced by Blunt Force with Different Heights / 法医学杂志

OBJECTIVES@#To explore the differences in the repair process of skin and skeletal muscle after contusion caused by blunt force attack with different heights.@*METHODS@#Three degrees of contusion were performed on SD rats' right hind limbs by a designed free-dropping device falling from 15, 30 and 50 cm heights, which as a main consideration factor for degree of injury. The repair process of skin and skeletal muscle at 6 h, 24 h, 3 d, 7 d and 13 d after contusion were observed using routine histological methods.@*RESULTS@#Hematoma within skin and/or muscle was found in the rats' hind limbs after contusion with three different heights. The repair processes were similar at 24 h after contusion. However, with the increase of height, the display degree was more obvious. At 3 d after contusion, the RBC of the hemorrhagic region would be decomposed and elapsed in 15 cm contusion group, but for 30 cm contusion group, it delayed to 7 d. At 13 d after contusion, the similar result was found in 15 cm and 30 cm contusion groups, in contrast, the 50 cm contusion group was still in the proliferative phase.@*CONCLUSIONS@#With the increase of height, the occurring rate of hematoma within skin and muscle at the same time increases, and the more serious histological appearance after contusion, including inflammation and proliferation, the longer healing process are observed. According to the results of present study and considering forensic application, the contusion model with 50 cm height （2.58 J/cm²） is recommended as the experimental animal model for the future study of wound age estimation on contusion.

Comparative kinematic gait analysis in young and old Beagle dogs

Age-related involution in dogs involves loss of muscle mass and changes in connective tissue and articular cartilage. The aim of this study was to examine whether an age-related influence on joint mobility can be detected in the absence of disease. Five young (mean age 2.0 years) and five old (mean age 10.4 years) healthy and sound Beagle dogs underwent computer-assisted gait analysis during locomotion on a treadmill. Shoulder, elbow, carpal, hip, stifle, and tarsal joint angles including joint angle progression curves, minimum and maximum joint angles, and range of motion (ROM) in degrees were analyzed. The old group had a smaller maximum joint angle (p = 0.037) and ROM (p = 0.037) of the carpal joint; there were similar tendencies in the shoulder, elbow, and carpal joints. Descriptive analysis of the progression curves revealed less flexion and extension of the forelimb joints. The results indicate restricted joint mobility of the forelimb in old dogs, primarily of the carpal joint. Results in the joints of the hindlimb were inconsistent, and the contrasting alterations may be due to a compensatory mechanism. As most alterations were found in the distal joints, these should receive particular attention when examining elderly dogs.

Effects of N-acetylcysteine and pentoxifylline on remote lung injury in a rat model of hind-limb ischemia/reperfusion injury / Efeitos da N-acetilcisteína e pentoxifilina na lesão pulmonar remota em um modelo de lesão de isquemia/reperfusão de membro posterior em ratos

Objective : To investigate the effects of N-acetylcysteine (NAC) and pentoxifylline in a model of remote organ injury after hind-limb ischemia/reperfusion (I/R) in rats, the lungs being the remote organ system. Methods : Thirty-five male Wistar rats were assigned to one of five conditions (n = 7/group), as follows: sham operation (control group); hind-limb ischemia, induced by clamping the left femoral artery, for 2 h, followed by 24 h of reperfusion (I/R group); and hind-limb ischemia, as above, followed by intraperitoneal injection (prior to reperfusion) of 150 mg/kg of NAC (I/R+NAC group), 40 mg/kg of pentoxifylline (I/R+PTX group), or both (I/R+NAC+PTX group). At the end of the trial, lung tissues were removed for histological analysis and assessment of oxidative stress. Results : In comparison with the rats in the other groups, those in the I/R group showed lower superoxide dismutase activity and glutathione levels, together with higher malondialdehyde levels and lung injury scores (p < 0.05 for all). Interstitial inflammatory cell infiltration of the lungs was also markedly greater in the I/R group than in the other groups. In addition, I/R group rats showed various signs of interstitial edema and hemorrhage. In the I/R+NAC, I/R+PTX, and I/R+NAC+PTX groups, superoxide dismutase activity, glutathione levels, malondialdehyde levels, and lung injury scores were preserved (p < 0.05 for all). The differences between the administration of NAC or pentoxifylline alone and the administration of the two together were not significant for any of those parameters (p > 0.05 for all). Conclusions : Our results suggest that NAC and pentoxifylline both protect lung tissue from the effects of skeletal muscle I/R. However, their combined use does not appear to increase the level of that protection.

Objetivo : Investigar os efeitos da N-acetilcisteína (NAC) e pentoxifilina em um modelo de lesão pulmonar remota após isquemia/reperfusão (I/R) de membro posterior em ratos. Métodos : Trinta e cinco ratos Wistar machos foram divididos em cinco grupos (n = 7/grupo), cada qual submetido ao seguinte: operação simulada (grupo controle); isquemia de membro posterior, induzida por pinçamento da artéria femoral esquerda por 2 h, seguida por de 24 h de reperfusão (grupo I/R); e isquemia de membro posterior, como descrito acima, seguida de injeção intraperitoneal (antes da reperfusão) de 150 mg/kg de NAC (grupo I/R+NAC), 40 mg/kg de pentoxifilina (grupo I/R+PTX) ou ambas (grupo I/R+NAC+PTX). Ao final do experimento, tecidos pulmonares foram removidos para análise histológica e avaliação do estresse oxidativo. Resultados : Comparados aos ratos dos outros grupos, os do grupo I/R apresentaram menor atividade de superóxido dismutase e menores níveis de glutationa, além de maiores níveis de malondialdeído e maiores escores de lesão pulmonar (p < 0,05 para todos). Infiltração celular inflamatória intersticial dos pulmões também foi bem maior no grupo I/R do que nos outros grupos. Além disso, os ratos do grupo I/R apresentaram vários sinais de edema intersticial e hemorragia. Nos grupos I/R+NAC, I/R+PTX e I/R+NAC+PTX, a atividade de superóxido dismutase, níveis de glutationa, níveis de malondialdeído e escores de lesão pulmonar foram preservados (p < 0,05 para todos). As diferenças entre a administração de NAC ou pentoxifilina isoladamente e a das duas combinadas não foi significativa para nenhum desses parâmetros (p > 0,05 para todos). Conclusões : Nossos resultados sugerem que tanto NAC quanto pentoxifilina protegem o tecido pulmonar dos efeitos de I/R de músculo esquelético. Entretanto, seu uso combinado não parece aumentar o nível dessa proteção.

Directing Human Embryonic Stem Cells towards Functional Endothelial Cells Easily and without Purification

Hemangioblasts or blood islands only arise in early development thereby the sources to obtain these bi-potential cells are limited. While previous studies have isolated both lineages in vitro through the hemangioblast, derivation efficiency was rather low due to cellular damage attributed by enzyme usage and fluorescent activated cell sorting (FACS). This study focused on avoiding the use of damaging factors in the derivation of endothelial cells (ECs). Single cell H9-human embryonic stem cells (hESCs) were obtained by using a mild dissociation protocol then human embryoid body (hEB) formation was performed under hemangioblast differentiation conditions. The hEBs were subjected to a two-stage cytokine treatment procedure. Subsequent culture of the adhesive cells in day 4 hEBs gave arise to a seemingly pure population of ECs. The hESC-derived ECs were characterized by identifying signature endothelial gene and protein markers as well as testing for in vitro functionality. Furthermore, in vivo functionality was also confirmed by transplanting the cells in hindlimb ischemic murine models. We demonstrate that the genetic change required for EC derivation precedes blast colony formation. Furthermore, cell damage was prevented by abating enzyme usage and FACS, resulting in a high yield of ECs upon adhesion. Under this method, confluent cultures of ECs were obtainable 4 days after hEB formation which is significantly faster than previous protocols.